All-Optical Volumetric Physiology for Connectomics in Dense Neuronal Structures
Chiao Huang,
Chu-Yi Tai,
Kai-Ping Yang,
Wei-Kun Chang,
Kuo-Jen Hsu,
Ching-Chun Hsiao,
Shun-Chi Wu,
Yen-Yin Lin,
Ann-Shyn Chiang,
Shi-Wei Chu
Affiliations
Chiao Huang
Department of Physics, National Taiwan University, 1, Sec 4, Roosevelt Road, Taipei 10617, Taiwan
Chu-Yi Tai
Institute of Biotechnology, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan
Kai-Ping Yang
Department of Physics, National Taiwan University, 1, Sec 4, Roosevelt Road, Taipei 10617, Taiwan
Wei-Kun Chang
Brain Research Center, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan
Kuo-Jen Hsu
Department of Physics, National Taiwan University, 1, Sec 4, Roosevelt Road, Taipei 10617, Taiwan; Brain Research Center, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan
Ching-Chun Hsiao
Department of Engineering and System Science, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan
Shun-Chi Wu
Department of Engineering and System Science, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan
Yen-Yin Lin
Brain Research Center, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan; Corresponding author
Ann-Shyn Chiang
Institute of Biotechnology, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan; Brain Research Center, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan; Institute of Systems Neuroscience, National Tsing Hua University, 101, Sec 2, Guangfu Road, Hsinchu 30013, Taiwan; Department of Biomedical Science and Environmental Biology, Kaohsiung Medical University, Kaohsiung 80780, Taiwan; Graduate Institute of Clinical Medical Science, China Medical University, Taichung 40402, Taiwan; Institute of Molecular and Genomic Medicine, National Health Research Institutes, Zhunan, Miaoli 35053, Taiwan; Kavli Institute for Brain and Mind, University of California, San Diego, CA 92161, USA; Corresponding author
Shi-Wei Chu
Department of Physics, National Taiwan University, 1, Sec 4, Roosevelt Road, Taipei 10617, Taiwan; Molecular Imaging Center, National Taiwan University, 1, Sec 4, Roosevelt Road, Taipei 10617, Taiwan; Corresponding author
Summary: All-optical physiology (AOP) manipulates and reports neuronal activities with light, allowing for interrogation of neuronal functional connections with high spatiotemporal resolution. However, contemporary high-speed AOP platforms are limited to single-depth or discrete multi-plane recordings that are not suitable for studying functional connections among densely packed small neurons, such as neurons in Drosophila brains. Here, we constructed a 3D AOP platform by incorporating single-photon point stimulation and two-photon high-speed volumetric recordings with a tunable acoustic gradient-index (TAG) lens. We demonstrated the platform effectiveness by studying the anterior visual pathway (AVP) of Drosophila. We achieved functional observation of spatiotemporal coding and the strengths of calcium-sensitive connections between anterior optic tubercle (AOTU) sub-compartments and >70 tightly assembled 2-μm bulb (BU) microglomeruli in 3D coordinates with a single trial. Our work aids the establishment of in vivo 3D functional connectomes in neuron-dense brain areas. : Neuroscience; Techniques in Neuroscience; Biological Sciences Research Methodologies; Biological Sciences Tools Subject Areas: Neuroscience, Techniques in Neuroscience, Biological Sciences Research Methodologies, Biological Sciences Tools